WO2009051844A1 - Analogues synthétiques de peptides de régénération neurale - Google Patents

Analogues synthétiques de peptides de régénération neurale Download PDF

Info

Publication number
WO2009051844A1
WO2009051844A1 PCT/US2008/011951 US2008011951W WO2009051844A1 WO 2009051844 A1 WO2009051844 A1 WO 2009051844A1 US 2008011951 W US2008011951 W US 2008011951W WO 2009051844 A1 WO2009051844 A1 WO 2009051844A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
nrp
compound
nrp compound
peptide
Prior art date
Application number
PCT/US2008/011951
Other languages
English (en)
Inventor
Paul William Richard Harris
Margaret Anne Brimble
Frank Sieg
Original Assignee
Neuren Pharmaceuticals Limited
Neuren Pharmaceuticals, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to EP08840727.5A priority Critical patent/EP2212349B1/fr
Priority to SI200831077T priority patent/SI2212349T1/sl
Priority to ES08840727T priority patent/ES2433568T3/es
Priority to DK08840727.5T priority patent/DK2212349T3/da
Priority to JP2010529981A priority patent/JP5425086B2/ja
Priority to AU2008314565A priority patent/AU2008314565B2/en
Priority to PL08840727T priority patent/PL2212349T3/pl
Priority to NZ585363A priority patent/NZ585363A/en
Application filed by Neuren Pharmaceuticals Limited, Neuren Pharmaceuticals, Inc. filed Critical Neuren Pharmaceuticals Limited
Publication of WO2009051844A1 publication Critical patent/WO2009051844A1/fr
Priority to US12/754,937 priority patent/US8309684B2/en
Priority to US13/651,884 priority patent/US9040485B2/en
Priority to HRP20131016AT priority patent/HRP20131016T1/hr
Priority to US14/701,991 priority patent/US9650418B2/en
Priority to US15/486,734 priority patent/US20170218023A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/48—Nerve growth factor [NGF]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/28—Neurological disorders
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/28—Neurological disorders
    • G01N2800/2835—Movement disorders, e.g. Parkinson, Huntington, Tourette
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/28—Neurological disorders
    • G01N2800/285—Demyelinating diseases; Multipel sclerosis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/28—Neurological disorders
    • G01N2800/2878—Muscular dystrophy
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/28—Neurological disorders
    • G01N2800/2878—Muscular dystrophy
    • G01N2800/2892—Myotonic dystrophy

Definitions

  • This invention relates to synthetic analogues of peptides that have neural regeneration, migration, proliferation, differentiation and/or axonal outgrowth properties. These peptides are termed "Neural Regeneration Peptides” or "NRPs.” In particular, this invention relates to analogues of relatively small peptides that have one or more biological properties of NRPs.
  • NRPs Neural regeneration peptides
  • These functions include neural survival, neural proliferation, neuronal outgrowth, neural migration and neuronal differentiation.
  • Several NRPs have been previously described, and include those disclosed in US Patent Application Nos: 10/225,838 and 10/976,699, PCT/US02/026782, PCT/US2004/036203, PCT/US2006017534 and PCT/US2006026994.
  • 10/225,838 and 10/976,699 PCT/US02/026782
  • PCT/US2004/036203 PCT/US2006017534
  • PCT/US2006026994 PCT/US2006026994.
  • NRPs described to date have desirable pharmacodynamic properties and promote neural regeneration, migration, proliferation, differentiation and/or axonal outgrowth.
  • synthetic NRP analogues that also have improved pharmacokinetic properties.
  • synthetic molecules or modified peptides that have desirable pharmacodynamic properties similar to those of NRPs but also have improved pharmacokinetic properties and/or are chemically stable.
  • Certain aspects of this invention include novel synthetic NRP analogue molecules that can be used to treat disorders of the nervous system or other systems in which NPRs is effective. Another aspect is to provide therapies for disorders of cellular degeneration and death, including certain nervous system disorders.
  • synthetic analogues of NRPs can be used to treat adverse effects of amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), oxidative stress (e.g., Huntington's disease) or peripheral neuropathy (PN).
  • ALS amyotrophic lateral sclerosis
  • MS multiple sclerosis
  • oxidative stress e.g., Huntington's disease
  • PN peripheral neuropathy
  • An additional aspect of this invention is the production of NRP analogues with improved stability.
  • NRP compound analogue of NRP
  • SEQ ID NO: an amino acid sequence
  • an “analogue of NRP” may be characterized herein as having a particular amino acid sequence, a particular 2-dimensional representation of the structure, but it is understood that the actual molecule claimed has other features, including 3- dimensional structure, mobility about certain bonds and other properties of the molecule as a whole. It is the molecules themselves and their properties as a whole that are the subjects of this invention.
  • NRP neuropeptide
  • analogues of relatively short NRPs are provided that can have increased stability, due at least in part to decreased enzymatic degradation.
  • NRP analogues are provided having modified amino acids.
  • NRP analogues are provided that have non-amino acid substituents replacing amino acids.
  • FIG. 1 depicts a graph of neuroprotective effects of two NRPs, SEQ ID NO: 5 of this invention and SEQ ID NO:1 in cell cultures exposed to the neurotoxin 3-NP.
  • FIG. 2 depicts a graph of results of studies of neuroprotective effective of SEQ ID NO: 1 in which the peptide was stored at either -2O 0 C or -4 0 C.
  • FIG. 3 depicts a graph of results of studies of neuroprotective effects of SEQ ID NO: 5 of this invention in which the peptide was stored at either -2O 0 C or -4 0 C.
  • FIG. 4 depicts a graph of results of an enlarged study of neuroprotective effects of SEQ ID NO:5 of this invention and SEQ ID NO: 1 in cell cultures treated with the neurotoxin 3-NP, similar to those shown in FIG. 1.
  • FIG. 5 depicts a graph showing significant long-term effects of sequence
  • REGRRDAPGRAGG (SEQ ID NO: 12) of this invention to decrease the severity of motoric impairment in animals with EAE, when the synthetic NRP was administered at the peak of the disease.
  • Score 1 is the lowest score and implies a flaccid tail only, while the higher scores imply weakness (score 2) or complete paralysis of the hind legs (score 3).
  • Kruskal-Wallis-test was used for statistical analysis; ** p ⁇ 0.01 versus treatment day 1 score (data expressed as mean ⁇ SEM).
  • FIG. 6 depicts a graph of results of beam walking scores for rats with peripheral neuropathy induced by pyridoxine (800 mg/kg/day) and treated with either vehicle or SEQ ID NO:5 of this invention at two different doses.
  • FIG. 7 depicts a graph of results of beam walking scores for rats with peripheral neuropathy induced by pyridoxine (1200 mg/kg) and treated with either vehicle or SEQ ID NO:5 of this invention.
  • FIG. 8 depicts graphs of results of studies of longevity of mice with a murine model of amytrophic lateral sclerosis (ALS) and the effects of two different doses of a synthetic NRP of this invention, SEQ ID NO:5.
  • ALS amytrophic lateral sclerosis
  • NRP compounds are provided that have a sequence of a native peptide.
  • one such NRP is an 1 1 amino acid long (1 1-mer) peptide having the following sequence. NH 2 -G 1 RRAAPGRAGG 1 '-NH 2 SEQ ID NO: 1
  • NRP N-terminal hydroxyl residues
  • Synthetic analogues of NPRs are provided that can have one or more of the following types of modifications: (1) stabilization of ⁇ -turns, (2) replacement of glycine residues, (3) replacement of the N-terminal glycine residue and/or (4) cyclization.
  • sequence APGR (SEQ ID NO: 2)
  • the alanine or glycine can be replaced with aminoisobutyric acid (Aib).
  • substitution of the alanine with Aib produces the following analogue: NH 2 -G 1 RRA-AIb-PGRAGG 1 '-NH 2 SEQ ID NO:4
  • RAGG (SEQ ID NO: 3)
  • the alanine can be replaced with aminoisobutyric acid (Aib) to produce the analogue having the sequence : NH 2 -G 1 RRAAPGR-AIb-GG 1 '-NH 2 SEQ ID NO:5
  • SEQ ID NO:5 was: neuroprotective (see FIGs 1-4), stable under storage conditions (FIGs. 2 and 3), more neuroprotective than the unsubstituted NRP (FIGs. 1 and 4), peripheral neuropathy (FIGs. 6 and 7) and ALS (FIG. 8).
  • This NRP analogue was found to be neuroprotective in in vitro model of neural toxicity induced by 3-NP.
  • Truncation of the G 1 at the N terminus can result in loss of biological activity. Replacement of the G 1 with an acetyl group can restore biological activity.
  • the resulting NRP analogue is acetylated providing a peptide having the following sequence:
  • the secondary structure of a peptide can be affected by the presence of D-amino acids replacing one or more L-amino acids (naturally occurring).
  • Replacement of the third amino acid from the N-terminus produces a compound having the following sequence: NH 2 -GR (D-Arg) AAPGRAGG-NH 2 SEQ ID NO:8
  • This NRP analogue was found to be neuroprotective in the in vitro model of neural toxicity induced by 3 -NP.
  • Cyclization Synthesis of a cyclic peptide mimetic of SEQ ID NO:1 can be carried out.
  • One method involves adding a cysteine residue to each end of the sequence, and then oxidizing the resultant product to produce a cyclic disulfide having the following sequence:
  • both the N and C terminal glycine residues can be replaced with a cysteine residue and oxidized similarly as above, producing an analogue having the following sequence.
  • Direct cyclization of the C terminal residue to the N terminal residue can be accomplished by creating an amide bond to produce a peptide having the following sequence.
  • synthetic production of the polypeptides of the invention may be according to the solid-phase synthetic method described by Goodman M. (ed.), "Synthesis of Peptides and Peptidomimetics” in Methods of organic chemistry (Houben-Weyl) (Workbench Edition, E22a,b,c,d,e; 2004; Georg Thieme Verlag, Stuttgart, New York)., herein expressly incorporated fully by reference.
  • This technique is well understood and is a common method for preparation of peptides. The general concept of this method depends on attachment of the first amino acid of the chain to a solid polymer by a covalent bond.
  • Amino acids may be attached to any suitable polymer as a resin.
  • the resin must contain a functional group to which the first protected amino acid can be firmly linked by a covalent bond.
  • Various polymers are suitable for this purpose, such as cellulose, polyvinyl alcohol, polymethylmethacrylate and polystyrene. Suitable resins are commercially available and well known to those of skill in the art.
  • protective groups usable in such synthesis include tert- butyloxycarbonyl (BOC), benzyl (BzI), t-amyloxycarbonyl (Aoc), tosyl (Tos), o-bromo- phenylmethoxycarbonyl (BrZ), 2,6-dichlorobenzyl (BzICl 2 ), and phenylmethoxycarbonyl (Z or CBZ). Additional protective groups are identified in Goodman, cited above, as well as in McOmie JFW: Protective Groups in Organic Chemistry, Plenum Press, New York, 1973, both references expressly incorporated fully herein by reference.
  • General procedures for preparing peptides of this invention involve initially attaching a carboxyl-terminal protected amino acid to the resin. After attachment the resin is filtered, washed and the protecting group on the alpha-amino group of the carboxyl-terminal amino acid is removed. The removal of this protecting group must take place, of course, without breaking the bond between that amino acid and the resin. The next amino, and if necessary, side chain protected amino acid, is then coupled to the free amino group of the amino acid on the resin. This coupling takes place by the formation of an amide bond between the free carboxyl group of the second amino acid and the amino group of the first amino acid attached to the resin. This sequence of events is repeated with successive amino acids until all amino acids are attached to the resin.
  • the protected peptide is cleaved from the resin and the protecting groups removed to reveal the desired peptide.
  • the cleavage techniques used to separate the peptide from the resin and to remove the protecting groups depend upon the selection of resin and protecting groups and are known to those familiar with the art of peptide synthesis.
  • Peptides may be cyclized by the formation of a disulfide bond between two cysteine residues. Methods for the formation of such bonds are well known and include such methods as those described in G. A. Grant (Ed.) Synthetic Peptides A User's Guide 2 nd Ed., Oxford University Press, 2002, W. C. Chan and P. D. White (Eds.) Fmoc Solid Phase Synthesis A Practical Approach, Oxford University Press, 2000 and references therein.
  • peptides of the invention may also be synthesized using standard solution peptide synthesis methodologies, involving either stepwise or block coupling of amino acids or peptide fragments using chemical or enzymatic methods of amide bond formation (see, e.g. H. D. Jakubke in The Peptides, Analysis, Synthesis, Biology, Academic Press, New York, 1987, p. 103- 165; J. D. Glass, ibid., pp.
  • NRP analogues of this invention can be used to treat neurological disorders.
  • NPRs have been unexpectedly effective in treating neural degeneration associated with autoimmune disorders of the brain, including EAE and multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and toxic injury to neural cells.
  • MS multiple sclerosis
  • ALS amyotrophic lateral sclerosis
  • NRP Analogues Disorders and conditions in which NRP compounds of this invention can be of benefit include the following.
  • Nervous system conditions treatable with NRP analogues include infections of the central nervous system including bacterial, fungal, spirochetal, parasitic and sarcoid including pyrogenic infections, acute bacterial meningitis or leptomeningitis.
  • Cerebrovascular diseases include stroke, ischemic stroke, hypoxia/ischemia, atherosclerotic thrombosis, lacunes, embolism, hypertensive haemorrhage, ruptured aneurysms, vascular malformations, transient ischemic attacks, intracranial haemorrhage, spontaneous subarachnoid haemorrhage, hypertensive encephalopathy, inflammatory diseases of the brain arteries, decreased perfusion caused by, for example, cardiac insufficiency (possibly resulting from coronary bypass surgery) and other forms of cerebrovascular disease.
  • Craniocerebral traumas include basal skull fractures and cranial nerve injuries, carotid- cavernous fistula, pneumocephalus, aerocele andrhinorrhea, cerebral contusion, traumatic intracerebral haemorrhage, traumatic brain injury, penetrating traumatic brain injury and acute brain swelling in children.
  • Demyelinating diseases include neuromyelitis optica, acute disseminated encephalomyelitis, acute and subacute necrotizing haemorrhagic encephalitis, diffuse cerebral sclerosis of Schilder and multiple sclerosis in conjunction with peripheral neuropathy.
  • Degenerative diseases of the nervous system including syndrome of one or more of progressive dementia, diffuse cerebral atrophy, diffuse cortical atrophy of the non-Alzheimer type, Lewy body dementia, Pick's disease, fronto-temporal dementia, thalamic degeneration, non-Huntingtonian types of Chorea and dementia, cortico-spinal degeneration (Jakob), the dementia-Parkinson-amyotrophic lateral sclerosis complex (Guamanina and others) and amyotrophic lateral sclerosis (ALS).
  • progressive dementia diffuse cerebral atrophy
  • diffuse cortical atrophy of the non-Alzheimer type Lewy body dementia
  • Pick's disease fronto-temporal dementia
  • thalamic degeneration non-Huntingtonian types of Chorea and dementia
  • cortico-spinal degeneration cortico-spinal degeneration
  • the dementia-Parkinson-amyotrophic lateral sclerosis complex (Guamanina and others)
  • Peripheral neuropathies are common and disabling conditions characterised by damage to or loss of peripheral neurons.
  • peripheral neuropathy There are more than 100 types of peripheral neuropathy, each with its own characteristic set of symptoms, pattern of development, and prognosis.
  • Peripheral neuropathy may be either inherited or acquired. Inherited forms of peripheral neuropathy can be caused by genetic mutations.
  • Some types of peripheral neuropathy and features common to them are shown below in Table 1. Table 1 above shows comparisons between pyridoxine-, streptozotin- and chemotherapy- induced peripheral neuropathy and diabetic peripheral neuropathy.
  • Acquired peripheral neuropathy may result from: physical injury (trauma) to a nerve, tumors, toxins (including chemotherapy), autoimmune responses, nutritional deficiencies, alcoholism, vascular and metabolic disorders (e.g.
  • the HIV-associated peripheral neuropathy is a common side effect of drugs targeting the reverse transcriptase of the HIV virus.
  • the symptoms of peripheral neuropathy can vary from temporary numbness, tingling, and pricking sensations, sensitivity to touch or muscle weakness, to more extreme symptoms such as burning pain, muscle wasting, paralysis, organ or gland dysfunction.
  • rat models of peripheral neuropathy enables the examination of neurological abnormalities induced by pyridoxine. For instance, 1,200 or 800mg/kg/day of pyridoxine administered to rats for 5-10 days results in necrosis of sensory neurons, especially affecting large diameter neurons in the sciatic nerve and the dorsal root ganglion (Xu et al., Neurology 39: 1077-
  • Pyridoxine-induced peripheral neuropathy in animals is a recognized system for studying effects of therapeutic agents.
  • this system is predictive of effects of such agents on peripheral neuropathy in human beings.
  • metabolic disorders of the nervous system including metabolic diseases presenting as a syndrome comprising one or more of confusion, stupor or coma-ischemia-hypoxia, hypoglycaemia, hyperglycemia, hypercapnia, hepatic failure and Reye syndrome, metabolic diseases presenting as a progressive extrapyramidal syndrome, metabolic diseases presenting as cerebellar ataxia, hyperthermia, celiac-sprue disease, metabolic diseases causing psychosis or dementia including Cushing disease and steroid encephalopathy, thyroid psychosis and hypothyroidism and pancreatic encephalopathy.
  • An example of a metabolic disorder that can result in neuropathy is pyridoxine excess described more fully below.
  • disorders of the nervous system due to drugs and other chemical agents include opiates and synthetic analgesics, sedative hypnotic drugs, stimulants, psychoactive drugs, bacterial toxins, plant poisons, venomous bites and stings, heavy metals, industrial toxins, anti-neoplastic and immunosuppressive agents, thalidomide, aminoglycoside antibiotics (ototoxicity) and penicillin derivatives (seizures), cardioprotective agents (beta-blockers, digitalis derivatives and amiodarone).
  • compositions and methods of the invention can find use in the treatment of human neural injury and disease. Still more generally, the compositions and methods of the invention find use in the treatment of human patients suffering from neural damage as the result of acute brain injury, including but not limited to diffuse axonal injury, perinatal hypoxic-ischemic injury, traumatic brain injury, stroke, ischemic infarction, embolism, and hypertensive haemorrhage; exposure to CNS toxins, infections of the central nervous system, such as, bacterial meningitis; metabolic diseases such as those involving hypoxic-ischemic encephalopathy, peripheral neuropathy, and glycogen storage diseases; or from chronic neural injury or neurodegenerative disease, including but not limited to Multiple Sclerosis, Lewy Body Dementia, Alzheimer's disease, Parkinson's disease and Huntington's disease.
  • Patient's suffering from such diseases or injuries may benefit greatly by a treatment protocol able to initiate neuronal proliferation and migration, as well as neurite outgrowth.
  • the invention has application in the induction of neuronal and neuroblast migration into areas of damage following insult in the form of trauma, toxin exposure, asphyxia or hypoxia-ischemia.
  • NRP analogues can be used via direct administration to the patient.
  • An NRP analogue can be administered as part of a medicament or pharmaceutical preparation. This can involve combining an NRP analogue with any pharmaceutically appropriate carrier, adjuvant or excipient. Additionally an NRP analogue can be used with other non-NRP neuroprotective, proliferative, or other agent. The selection of the carrier, adjuvant or excipient can depend upon the route of administration to be employed.
  • the administration route can vary widely to suit a particular condition.
  • An NRP analogue may be administered in different ways: intraperitoneally, intravenously or intracerebroventricularly.
  • the peripheral application may be a route of choice because then there is no direct interference with the central nervous system. Any peripheral route of administration known in the art can be employed. These can include parenteral routes for example injection into the peripheral circulation, subcutaneous, intraorbital, ophthalmic, intraspinal, intracisternal, topical, infusion (using eg.
  • compositions can be formulated for parenteral administration to humans or other mammals in therapeutically effective amounts (eg. amounts which eliminate or reduce the patient's pathological condition) to provide therapy for the neurological diseases described above.
  • One route of administration includes subcutaneous injection (e.g., dissolved in 0.9% sodium chloride) and oral administration (e.g., in a capsule). It will also be appreciated that it may on occasion be desirable to directly administer NRP analogue to the CNS of the patient by any appropriate route of administration. Examples include administration by lateral cerebroventricular injection or through a surgically inserted shunt into the lateral cerebral ventricle of the brain of the patient, into the cerebrospinal fluid or directly into an affected portion of a patient's brain.
  • methods for treating brain damage comprise administering one or more NRP analogues in a dose range of from about O.Ol ⁇ g/kg body weight to about 100 ⁇ g/kg body weight. In other embodiments, a dose of 1 ⁇ g/kg body weight to about 10 ⁇ g/kg body weight can be useful. In further embodiments, a dose of an NRP can be in the range of about 0.01 ⁇ g/kg body weight to about O.lmg/kg.
  • an effective amount of an NRP analogue to be administered is within the skill of one of ordinary skill in the art, and will be routine to those persons skilled in the art.
  • the amount of an NRP analogue to be used can be estimated by in vitro studies using an assay system as described herein. The final amount of an NRP analogue to be administered will be dependent upon the route of administration, upon the NRP analogue used and the nature of the neurological disorder or condition that is to be treated.
  • a suitable dose range may for example, be between about 0.0 l ⁇ g to about 15 ⁇ g per lkg of body weight or in other embodiments, about 20 ⁇ g/kg to about 30 ⁇ g/kg body weight per day.
  • NRP analogue For inclusion in a medicament, NRP analogue can be directly synthesized by conventional methods such as the stepwise solid phase synthesis method of Merrifield et al, 1963 (J. Am. Chem. Soc. 15:2149-2154) or Goodman M. (ed.), "Synthesis of Peptides and Peptidomimetics” in Methods of organic chemistry (Houben-Weyl) (Workbench Edition, E22a,b,c,d,e; 2004; Georg Thieme Verlag, Stuttgart, New York), expressly incorporated herein fully by reference.
  • peptide synthesis can involve the use of commercially available peptide synthesizers such as the Applied Biosystems model 430A.
  • the total pharmaceutically effective amount of an NRP analogue administered parenterally per dose will be in a range that can be measured by a dose response curve.
  • an NRP analogue in the blood can be measured in body fluids of the mammal to be treated to determine dosing.
  • the amount of NRP analogue to be employed can be calculated on a molar basis based on these serum levels of the NRP analogue.
  • One method for determining appropriate dosing of the compound entails measuring NRP levels in a biological fluid such as a body or blood fluid. Measuring such levels can be done by any means, including RIA and ELISA. After measuring NRP analogue levels, the fluid is contacted with the compound using single or multiple doses. After this contacting step, the NRP analogue levels are re-measured in the fluid. If the fluid NRP analogue levels have fallen by an amount sufficient to produce the desired efficacy for which the molecule is to be administered, then the dose of the molecule can be adjusted to produce maximal efficacy. This method can be carried out in vitro or in vivo.
  • This method can be carried out in vivo, for example, after the fluid is extracted from a mammal and the NRP analogue levels measured, the compound herein is administered to the mammal using single or multiple doses (that is, the contacting step is achieved by administration to a mammal) and then the NRP analogue levels are re-measured from fluid extracted from the mammal.
  • NRP analogues are suitably administered by a sustained-release system.
  • sustained-release compositions include semi-permeable polymer matrices in the form of shaped articles, for example, films, or microcapsules.
  • Sustained-release matrices include polylactides (U.S. Pat. No. 3,773,919, EP 58,481), poly(2-hydroxyethyl methacrylate) (Langer et al, 1981), ethylene vinyl acetate (Langer et al, supra), or poly-D-(-)-3-hydroxybutyric acid (EP 133,988).
  • Sustained- release compositions also include a liposomally associated compound.
  • Liposomes containing the compound are prepared by methods known to those of skill in the art, as exemplified by DE 3,218,121 ; Hwang et al, 1980; EP 52,322; EP 36,676; EP 88,046; EP 143,949; EP 142,641; Japanese Pat. Appln. 83-1 18008, U.S. Pat. Nos. 4,485,045 and 4,544,545 and EP 102,324.
  • liposomes are of the small (from or about 200 to 800 Angstroms) unilamellar type in which the lipid content is greater than about 30 mol. percent cholesterol, the selected proportion being adjusted for the most efficacious therapy. All U.S. parents referred to herein, both supra and infra, are hereby expressly incorporated by reference in their entirety.
  • the compound can be formulated generally by mixing each at a desired degree of purity, in a unit dosage injectable form (solution, suspension, or emulsion), with a pharmaceutically, or parenterally, acceptable carrier, i.e., one that is non-toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation.
  • a pharmaceutically, or parenterally, acceptable carrier i.e., one that is non-toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation.
  • the formulation preferably does not include oxidizing agents and other compounds that are known to be deleterious to polypeptides. It can be appreciated that the above doses are not intended to be limiting. Other doses outside the above ranges can be determined by those with skill in the art.
  • formulations can be prepared by contacting a compound uniformly and intimately with liquid carriers or finely divided solid carriers or both. Then, if desired, the product can be shaped into the desired formulation.
  • the carrier is a parenteral carrier, alternatively, a solution that is isotonic with the blood of the recipient. Examples of such carrier vehicles include water, saline, Ringer's solution, a buffered solution, and dextrose solution. Non-aqueous vehicles such as fixed oils and ethyl oleate are also useful herein.
  • the carrier suitably contains minor amounts of additives such as substances that enhance isotonicity and chemical stability.
  • Such materials are desirably non-toxic to recipients at the dosages and concentrations employed, and include, by way of example only, buffers such as phosphate, citrate, succinate, acetic acid, and other organic acids or their salts; antioxidants such as ascorbic acid; low molecular weight (less than about ten residues) polypeptides, e.g., polyarginine or tripeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; glycine; amino acids such as glutamic acid, aspartic acid, histidine, or arginine; monosaccharides, disaccharides, and other carbohydrates including cellulose or its derivatives, glucose, mannose, trehalose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; counter-ions such as sodium; non-ionic surfactants such as polysorbates, polox
  • An NRP compound can be desirably formulated in such vehicles at a pH of from about 6.5 to about 8. Alternatively, the pH can be from about 4.5 to about 8. It will be understood that use of certain of the foregoing excipients, carriers, or stabilizers will result in the formation of salts of the compound.
  • the final preparation may be a stable liquid or lyophilized solid.
  • adjuvants can be used.
  • Typical adjuvants which may be incorporated into tablets, capsules, and the like are a binder such as acacia, corn starch, or gelatin; an excipient such as microcrystalline cellulose; a disintegrating agent like corn starch or alginic acid; a lubricant such as magnesium stearate; a sweetening agent such as sucrose or lactose; a flavoring agent such as peppermint, wintergreen, or cherry.
  • a binder such as acacia, corn starch, or gelatin
  • an excipient such as microcrystalline cellulose
  • a disintegrating agent like corn starch or alginic acid
  • a lubricant such as magnesium stearate
  • a sweetening agent such as sucrose or lactose
  • a flavoring agent such as peppermint, wintergreen, or cherry.
  • the dosage form is a capsule, in addition to the above materials, it may also contain a liquid carrier such as a
  • a syrup or elixir may contain the active compound, a sweetener such as sucrose, preservatives like propyl paraben, a coloring agent, and a flavoring agent such as cherry.
  • a sweetener such as sucrose
  • preservatives like propyl paraben a coloring agent
  • a flavoring agent such as cherry.
  • Sterile compositions for injection can be formulated according to conventional pharmaceutical practice. For example, dissolution or suspension of the active compound in a vehicle such as water or naturally occurring vegetable oil like sesame, peanut, or cottonseed oil or a synthetic fatty vehicle like ethyl oleate or the like may be desired. Buffers, preservatives, antioxidants, and the like can be incorporated according to accepted pharmaceutical practice.
  • an NRP analogue to be used for therapeutic administration may be sterile.
  • Sterility can be readily accomplished by filtration through sterile filtration membranes (e.g., membranes having pore size of about 0.2 micron).
  • Therapeutic compositions generally can be placed into a container having a sterile access port, for example an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.
  • an NRP analogue can be stored in unit or multi-dose containers, for example, sealed ampules or vials, as an aqueous solution or as a lyophilized formulation for reconstitution.
  • a lyophilized formulation 10-mL vials are filled with 5 ml of sterile-filtered 0.01% (w/v) aqueous solution of compound, and the resulting mixture is lyophilized.
  • the infusion solution can be prepared by reconstituting lyophilized compounds using bacteriostatic water or other suitable solvent.
  • kits may contain a predetermined amount of lyophilized NRP compound, a physiologically compatible solution for preparation of a dosage form, a mixing vial, a mixing device, and instructions for use.
  • kits can be manufactured and stored according to usual practices in the industry.
  • An NRP compound-containing composition may be administered by one or more of a variety of routes.
  • routes such as intravenous, intraperitoneal, intracerebral, intraventricular, inhalation, lavage, rectal, vaginal, transdermal, subcutaneous administration can be used.
  • Example 1 Effects of NRP Compounds on Survival and Proliferation of Cerebellar Microexplants
  • NRP Compound Preparation NRP compounds were provided by Auspep (Australia). The peptides were synthesized using standard solid-phase synthesis. The peptides were supplied with an amidated C-terminus, and were more than 95% pure as analyzed by MALDI-MS spectrum analysis. The peptides were stored lyophilized at -8O 0 C under argon in 0.5M sucrose or 0.5M trehalose until usage. They were reconstituted in PBS, alternatively in lOO ⁇ g/ml human transferrin/PBS or in other embodiments in 1 OO ⁇ g/ml BSA/PBS, in 0.5M sucrose or 0.5M trehalose.
  • Laminated cerebellar cortices of the two hemispheres were explanted from a P3, P4, P7 or P8 Wistar rat, cut into small pieces in GBSS with 0.65% D(+)glucose solution, and triturated by a 0.4 mm gauge needle and subsequently pressed through a 125 ⁇ m pore size sieve.
  • the obtained microexplants were centrifuged (60 X g) 2 times for a medium exchange into serum-free BSA- supplemented START V-medium (Biochrom). Finally, the microexplants were reconstituted in 500 ⁇ l STARTV-medium.
  • cerebellar microexplants were fixed after 2-3 days in culture after the following regime: microexplants were fixed by 2- minute, serial treatment with 0.4%; 1.2%; 3% paraformaldehyde, respectively, followed by a 5 min incubation in 4% paraformaldehyde/ 0.25% glutaraldehyde in 0.1 M sodium phosphate (pH 7.4).
  • Oxidative stress can result in neurodegeneration. This is one possible mechanism for the symptoms observed in human disorder with Huntington's disease.
  • the oxidative stress-producing toxin, 3-nitropropioinic acid (3-NP) has been previously shown to produce effects in experimental animals that mimic those effects seen in human beings with Huntington's disease.
  • studies of therapeutic drugs in experimental animals with 3-NP induced neurotoxicity are predictive of effects of those drugs in treating human beings with Huntington's disease or other disorders characterized by oxidative stress.
  • General methods for toxicological and drug administration experiments were designed such that 1/100 parts of toxin and neuroprotective drug were administered simultaneously to the freshly prepared cerebellar microexplants.
  • Glutamate was prepared as a 5OmM stock solution in MiIIiQ water while 5OmM 3-nitropropionic acid (3-NP) was pH-adjusted (pH 6.8-7.2) in MiIIiQ water.
  • concentrations of the oxidative stress inducing toxin, 3-nitropropionic acid (3-NP), and the excitotoxin, glutamate, in the assay were at concentrations of 0.5mM each.
  • Lyophilized NRP peptides were reconstituted in PBS or lOO ⁇ g/ml human transferrin as a lO ⁇ M stock solution. Subsequently, serial dilutions were made.
  • Cerebellar microexplants were cultivated for 48-72 hours at 34°C, 5% CO 2 in air and 100% humidity before they were fixed by increasing amounts of paraformaldehyde (0.4%, 1.2%, 3% and 4% - each treatment 2-3min).
  • cerebellar explants were exposed for 24 hours, at the beginning of culturing to dilutions of NRP (survival assay) or NRP and 0.1 ⁇ M BrdU (proliferation assay). Subsequently, 80% of the medium was changed without addition of new toxins and NRPs. The cerebellar cultures were fixed as described above after 3 days in vitro. The detection of the incorporated BrdU level is performed as described previously.
  • NRP analogues promoted increased neuronal survival in explants treated with 3-NP (see Examples 2, 3 and 4).
  • SEQ ID NO: 1 and SEQ ID NO:5 have the same amino acid sequence, with the exception that the Alanine (A) in position 9 of the amino acid sequence of SEQ ID NO: 1 was replaced by aminoisobutyric acid (Aib), thus producing a compound having the sequence shown in SEQ ID NO:5.
  • Aib replacement of Alanine can stabilize the beta-turn, and thus, produce a peptide having different 3-dimensional structure and different mobilities about certain bonds compared to the un-substituted peptide.
  • FIG. 1 depicts a graph of results of these studies. 3-NP alone produced profound loss of cells displaying neurites compared to vehicle-treated controls, indicating that the compound is indeed neurotoxic.
  • the peptide having the sequence SEQ ID NO: 1 at concentrations of 10 fM or 1 pM significantly decreased the neurotoxic effects of 3-NP (mean ⁇ SEM; p ⁇ 0.001 ; n 4).
  • SEQ ID NO:5 decreased 3-NP-induced neurotoxicity in a concentration dependent fashion, with a threshold of below about IfM and a maximal effect at a concentration of about 10OfM (mean ⁇ SEM; p ⁇ 0.001; n 4 each).
  • SEQ ID NO:5 is neuroprotective. This result means that SEQ ID NO: 5 can be a valuable therapeutic NRP compound for treating neural degeneration in humans suffering from neurological disorders. Further, because oxidative stress is known to induce neurodegeneration similar to the neurodegeneration observed in human beings with Huntingdon's disease, synthetic NRP compounds of this invention can be used to treat human beings with neurodegeneration caused by oxidative stress.
  • Example 3 Stability and Neuroprotective Effects of Un-Substituted NRP Compounds
  • SEQ ID NO: 1 we synthesized SEQ ID NO: 1 and then stored the peptide at temperatures of either -2O 0 C or -4 0 C for nine (9) weeks.
  • FIG. 2 depicts results of these studies. Cerebellar explants treated with 3-NP alone (light stippled bar) showed fewer cells exhibiting neurites compared to vehicle-treated control explants (open bar). SEQ ID NO: 1 that had been stored at -2O 0 C exhibited neuroprotective effects at all concentrations tested (from 10 '13 M; 100 fM to 10 10 M; 10OpM), with statistically significant effects observed at concentrations of 10 " " M and 10 " '° M.
  • FIG. 3 depicts a graph of these studies. Cerebellar explants were treated with vehicle alone
  • SEQ ID NO:5 When stored at -2O 0 C, SEQ ID NO:5 produced neuroprotective effects similar to those found for
  • SEQ ID NO: 1 that had been stored at -2O 0 C, as shown in Example 3 and depicted in FIG. 2.
  • SEQ ID NO:5 retained its neuroprotective effect.
  • the degree of neuroprotection provided by SEQ ID NO:5 after storage at -4 0 C was not statistically different from the degree of neuroprotection provided after storage at -2O 0 C.
  • This result was completely unexpected based on studies of SEQ ID NO:1 shown above.
  • the increased stability of the NPR having SEQ ID NO:5 means that this compound will be more suitable for storage and transportation under commonly used conditions.
  • Example S Neuroprotective Effects of Un-Substituted and Substituted NRPs
  • FIG. 4 depicts a graph of results of studies of cerebellar microexplants treated with the neurotoxin 3-NP, demonstrating neuroprotection by SEQ ID NO: 5 and SEQ ID NO: 1 in a study of 6 in each group.
  • SEQ ID NO: 1 and SEQ ID NO:5 protect neurons from dying after exposure to the neurotoxin 3-NP.
  • SEQ ID NO:5 and SEQ ID NO:1 can be useful therapeutic agents in treating human beings suffering from neurotoxicity.
  • NRPs have an impact on chronic inflammation in the CNS that leads to severe axonal damage and subsequent lesions (such as in multiple sclerosis; MS).
  • EAE experimental autoimmune encephalitis
  • MOG myelin oligodendrocyte glycoprotein
  • mice Female mice, 6-8 weeks-old, strain C57B1/6J weighing an average of 24 gms each were used. NRP preparation
  • SEQ ID NO: 30 (also known as SEQ ID NO: 30 disclosed in U.S. Patent Application Number 10/976,699), was supplied by Auspep (Australia).
  • the peptide SEQ ID NO: 12 was supplied with an amidated C- terminus, and was more than 95% pure as determined by MALDI-MS spectroscopy. HPLC. The sequence was confirmed by mass spectroscopy. The peptide was stored lyophilized at a temperature of -8O 0 C under argron gas until use. The peptide was reconstituted in PBS on the day of use.
  • a 200 ⁇ ul L of an emulsion containing 200 ⁇ ug of the encephalitogenic peptide MOG35-55 MEVGWYRSPFSRVVHLYRNGK SEQ ID NO: 13 was obtained from C S Bio Co. USA) in complete Freund adjuvant (Difco, Detroit, USA) containing 800 ⁇ ug Mycobacterium tuberculosis (Difco, Detroit, USA) was injected subcutaneously into one flank. Mice were immediately injected intraperitoneally with 400ng pertussis toxin (List Biological Laboratories, USA) and again 48 hours later.
  • mice were monitored daily and neurological impairment was scored on an arbitrary clinical score as follows: 0, no clinical sign; 1, flaccid tail; 2, hind limb weakness; 3, hind limb paralysis; 4, hind limb weakness and fore limb weakness; 5, paraplegia; 6, death.
  • NRP analogues of this invention can be useful therapeutic agents for treating peripheral neuropathy, we carried out a series of studies in rats with peripheral neuropathy induced by pyridoxine.
  • Rats were administered 400mg/kg pyridoxine chloride dissolved in sterile distilled water adjusted to neutral pH intraperitoneally (i.p) twice daily for 8 days and concurrently the peptide having the sequence SEQ ID NO:5 was administered i.p. for at total of 10 days. The rats were observed for a total of 29 days.
  • Experiment II Rats were administered a higher dose of pyridoxine (1200mg/kg/day) over a shorter period of time. Pyridoxine chloride dissolved in sterile distilled water adjusted to neutral pH was administered i.p. to the rats at a dose of 600mg/kg twice daily for 4 days. Concurrently the peptide having the sequence SEQ ID NO:5 was administered for 4 days. On day 5, rats were tested for motor deficits. Motor deficits after pyridoxine intoxication and effects of SEQ ID NO:5 were analyzed using a precision beam walk.
  • SEQ ID NO:5 showed highly significant and clinically substantial attenuation of motor deficits pyridoxine-induced peripheral neuropathy.
  • SEQ ID NO:5 was well tolerated.
  • the 4 ⁇ g/kg/day concentration worked equally well in the more chronic model of pyridoxine intoxication (800mg/kg/day for 8 days) and the acute pyridoxine intoxication induced by 1200mg/kg/day administered for 4 days.
  • SEQ ID NO:5 can be effective treatment for peripheral neuropathies in human beings.
  • Example 8 Effect of SEQ ID NO:5 on Animals With Amytrophic Lateral Sclerosis
  • mice were randomly allocated to receive either vehicle or SEQ ID NO:5 from the point of disease onset onwards. Disease onset in each treatment allocation group was not significantly different: 92-93 days.
  • SEQ ID NO:5 treatment (40 ⁇ g/kg, given I/day, i.p. (FIG. 8A) or 0.4 ⁇ g/kg, given I/day, i.p. (FIG. 8B)) was started at the day of onset of the disease.
  • FIGs. 8A and 8B show Kaplan- Meier survival probability curves for the SOD-I mutant (ALS) transgenic mice.
  • NRP analogues of this invention can be useful in treating acute as well as chronic neurodegenerative disorders including ALS, neurotoxicity, neurodegeneration associated with oxidative stress, autoimmune disorders, traumatic brain injury and other neurological diseases and conditions. Further, we conclude that NRP analogues can have beneficial therapeutic effects in situations where loss of neurological function is a symptom.
  • NRP analogue is tested for migration-inducing/chemoattractive activity on mouse neural stem cells in a haptotactic migration assay as described below.
  • Control wells of Transwell plates (Corning) with 12 ⁇ m pore size are coated in 1.5ml of the BSA/PBS vehicle. Remaining plates are coated using O. lng/ml of NRP analogue (prepared in PBS containing lOug/ml BSA).
  • Laminin (7 ⁇ g/ml) is used as extracellular matrix (ECM) coating for mouse primary stem cells.
  • ECM extracellular matrix
  • the matrix is incubated at 37 0 C; 5% CO 2 for 2hrs at room temperature.
  • the cells are seeded onto the inserts (30,000 cells per well). Plates are fixed at 1-2 days in vitro (DIV).
  • a 5ug/mL PDL/PLL mixture (in PBS) is used to coat inserts. Subsequently the inserts are rinsed with MiIIiQ water.
  • Inserts are discarded and wells fixed in successive dilutions of PFA (0.4, 1.2, 3 and 4%) for 3-5min in each dilution.
  • the wells are rinsed and stored in successive dilutions of PFA (0.4, 1.2, 3 and 4%) 3-5 min in each dilution.
  • the wells are rinsed and stored in PBS until counting. All cells that display neurite outgrowth and traveled to the bottom chamber are counted as migrating cells. Results
  • NRP analogues can induce neuronal cell migration, and that they each can be used to treat neurodegeneration associated with neural injury or disease.
  • NRP analogue is tested for migration-inducing/chemoattractive activity on mouse neural stem cells in a haptotactic migration assay in injury conditions, as described below.
  • Pl (postnatal day 1) Wistar or Sprague Dawley rats are sacrificed by decapitation. Cortical heminspheres are removed and collected into separate tubes containing 4ml DMEM - 1 cortex per tube. The tissue is mechanically triturated. Cells are transferred into medium using a sterile pipette and filtered through a lOOum cell strainer into a 50ml centrifuge tube. Each tube is stocked up to 50ml with DMEM. The tubes are centrifuged for 5mins at 350 x g at 22 0 C. The cells are resuspended in 40ml of DMEM + 10% FBS.
  • the cells are then seeded into a 12-well plate + 5nM ocadaic acid (to remove neurons by inducing apoptotic cell death) and incubated at 37°C/10% CO 2 for 24 hrs in a Boyden Chamber.
  • the medium + FBS is replaced after 1 day with fresh DMEM + 10% FBS.
  • the cell growth is monitored until confluency (14-18 days).
  • TGF ⁇ l transforming growth factor ⁇ l
  • lOng/ml TGF ⁇ l is administered to the astrocytic monolayer for 24hrs. Additionally, astrocytic cultures re mechanically injured by a scalpel (one scratch throughout the bottom of the well).
  • Inserts are discarded and wells fixed in successive dilutions of PFA (0.4, 1.2, 3 and 4%) for 3-5min in each dilution.
  • the wells are rinsed and stored in successive dilutions of PFA (0.4, 1.2, 3 and 4%) 3-5 min in each dilution.
  • the wells are rinsed and stored in PBS until counting. All cells that display neurite outgrowth and traveled to the bottom chamber are counted as migrating cells.
  • Migrated stem cell number of labelled cells re analysed after 24hrs by a fluorescence-based computerized imaging system (Discovery- 1).
  • NRP analogues stimulate more stem cells to migrate than vehicle-treated controls. We conclude that NRP analogue induces neuronal stem cell migration, and that NRP analogues can be useful to treat neurodegeneration associated with neural injury or disease.
  • Cisplatin neurotoxicity the relationship between dosage, time, and platinum concentration in neurologic tissues, and morphologic evidence of toxicity. J. Clin. Oncol. 10: 795-803.
  • Compounds and compositions of this invention find industrial use in many aspects of commerce, including pharmaceutical manufacturing, formulation, and sale.
  • Methods of using compounds and compositions of this invention find industrial use in medical fields of neurology, and in particular, for treatment of neurological disorders in animals and human beings.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Veterinary Medicine (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Biomedical Technology (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Hospice & Palliative Care (AREA)
  • Psychiatry (AREA)
  • Urology & Nephrology (AREA)
  • Vascular Medicine (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne, selon certains modes de réalisation, des composés synthétiques (analogues de NRP) de peptides nommés peptides de régénération neurale (NRP). Les analogues de NRP sont obtenus en substituant des acides aminés dans la séquence peptidique native, en modifiant des acides aminés chimiquement, en remplaçant des acides aminés avec des fonctions synthétiques, par stabilisation des virages β, par acétylation des résidus de glycine terminaux ou par cyclisation. Les analogues de NRP peuvent être utilisés pour traiter diverses maladies qui impliquent une dégénération des cellules neurales, notamment pour traiter des maladies du système nerveux, notamment la neuropathie périphérique, la sclérose en plaques, la neuropathie périphérique diabétique, la neurodégénération induite par neurotoxine et la sclérose latérale amyotrophique.
PCT/US2008/011951 2007-10-17 2008-10-17 Analogues synthétiques de peptides de régénération neurale WO2009051844A1 (fr)

Priority Applications (13)

Application Number Priority Date Filing Date Title
PL08840727T PL2212349T3 (pl) 2007-10-17 2008-10-17 Syntetyczne analogi peptydów regeneracji neuronów
ES08840727T ES2433568T3 (es) 2007-10-17 2008-10-17 Análogos sintéticos de péptidos de regeneración neuronal
DK08840727.5T DK2212349T3 (da) 2007-10-17 2008-10-17 Syntetiske analoger af nerveregenationspeptider
JP2010529981A JP5425086B2 (ja) 2007-10-17 2008-10-17 神経再生ペプチド化合物
AU2008314565A AU2008314565B2 (en) 2007-10-17 2008-10-17 Synthetic analogues of neural regeneration peptides
EP08840727.5A EP2212349B1 (fr) 2007-10-17 2008-10-17 Analogues synthétiques de peptides de régénération neurale
NZ585363A NZ585363A (en) 2007-10-17 2008-10-17 Synthetic analogues of neural regeneration peptides
SI200831077T SI2212349T1 (sl) 2007-10-17 2008-10-17 Sintetični analogi nevralnih regeneracijskih peptidov
US12/754,937 US8309684B2 (en) 2007-10-17 2010-04-06 Synthetic analogues of neural regeneration peptides
US13/651,884 US9040485B2 (en) 2007-10-17 2012-10-15 Synthetic analogues of neural regeneration peptides
HRP20131016AT HRP20131016T1 (hr) 2007-10-17 2013-10-25 Sintetiäśki analozi neuralnih regeneracijskih peptida
US14/701,991 US9650418B2 (en) 2007-10-17 2015-05-01 Synthetic analogues of neural regeneration peptides
US15/486,734 US20170218023A1 (en) 2007-10-17 2017-04-13 Synthetic analogues of neural regeneration peptides

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US99929207P 2007-10-17 2007-10-17
US60/999,292 2007-10-17
US99950307P 2007-10-18 2007-10-18
US60/999,503 2007-10-18

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US74593710A Continuation 2007-10-17 2010-04-06
US12/754,937 Continuation US8309684B2 (en) 2007-10-17 2010-04-06 Synthetic analogues of neural regeneration peptides

Publications (1)

Publication Number Publication Date
WO2009051844A1 true WO2009051844A1 (fr) 2009-04-23

Family

ID=40567715

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2008/011951 WO2009051844A1 (fr) 2007-10-17 2008-10-17 Analogues synthétiques de peptides de régénération neurale

Country Status (12)

Country Link
US (4) US8309684B2 (fr)
EP (1) EP2212349B1 (fr)
JP (1) JP5425086B2 (fr)
AU (1) AU2008314565B2 (fr)
CY (1) CY1114619T1 (fr)
DK (1) DK2212349T3 (fr)
ES (1) ES2433568T3 (fr)
HR (1) HRP20131016T1 (fr)
NZ (1) NZ585363A (fr)
PL (1) PL2212349T3 (fr)
PT (1) PT2212349E (fr)
WO (1) WO2009051844A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012102625A2 (fr) * 2011-01-28 2012-08-02 Curonz Holdings Company Limited Composition thérapeutique
WO2012105854A2 (fr) * 2011-01-31 2012-08-09 Curonz Holdings Company Limited Compositions pour administration oculaire
WO2014017927A1 (fr) 2012-07-27 2014-01-30 Curonz Holdings Company Limited Procédé de traitement d'une lésion du nerf optique, d'une ischémie ophtalmique ou d'une lésion de reperfusion ophtalmique
WO2015084188A2 (fr) 2013-12-04 2015-06-11 Curonz Holdings Company Limited Méthode de traitement ou de prévention de l'épilepsie ou des crises d'épilepsie faisant appel à un ou plusieurs peptides stimulant la régénération neuronale
EP2964332A4 (fr) * 2013-03-08 2016-09-14 Curonz Holdings Company Ltd Peptides de régénération neurale et leurs utilisations

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2212349B1 (fr) * 2007-10-17 2013-08-07 Curonz Holdings Company Limited Analogues synthétiques de peptides de régénération neurale
EP3895705A3 (fr) * 2013-12-20 2022-03-09 University of Manitoba Procédés et compositions pour le traitement de neuropathies périphériques
WO2015194970A2 (fr) * 2014-06-20 2015-12-23 Curonz Holdings Company Limited Méthode de traitement ou de prévention de la sclérose en plaques au moyen d'un ou de plusieurs peptides de régénération neuronale

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995032003A1 (fr) 1994-05-24 1995-11-30 Amgen Boulder Inc. Facteurs de croissance proches de l'insuline modifies
US20040180823A1 (en) * 2002-09-24 2004-09-16 Pasquale Elena B. Novel agents that modulate Eph receptor activity
US20050131212A1 (en) * 2001-08-24 2005-06-16 Neuren Pharmaceuticals Ltd. Neural regeneration peptides and methods for their use in treatment of brain damage

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003018754A2 (fr) * 2001-08-24 2003-03-06 Neuronz Limited Peptides de regeneration neurale et procedes d'utilisation de ces derniers dans le traitement des lesions cerebrales
WO2005042561A2 (fr) * 2003-10-31 2005-05-12 Neuren Pharmaceuticals Limited Regeneration neuronale de peptides et methodes d'utilisation dans le traitement des troubles cerebraux
JP4988711B2 (ja) * 2005-05-06 2012-08-01 クロンズ ホールディングズ カンパニー リミテッド 神経再生ペプチド及びそれを含む処方物
WO2007011595A2 (fr) * 2005-07-15 2007-01-25 Neuren Pharmaceuticals Limited Peptides de regeneration neurale et antioxydants protegeant les neurones de la degenerescence
EP2212349B1 (fr) * 2007-10-17 2013-08-07 Curonz Holdings Company Limited Analogues synthétiques de peptides de régénération neurale

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995032003A1 (fr) 1994-05-24 1995-11-30 Amgen Boulder Inc. Facteurs de croissance proches de l'insuline modifies
US20050131212A1 (en) * 2001-08-24 2005-06-16 Neuren Pharmaceuticals Ltd. Neural regeneration peptides and methods for their use in treatment of brain damage
US20040180823A1 (en) * 2002-09-24 2004-09-16 Pasquale Elena B. Novel agents that modulate Eph receptor activity

Non-Patent Citations (16)

* Cited by examiner, † Cited by third party
Title
BELL RH; HYE RJ: "Animal models of diabetes mellitus: physiology and pathology.", J. SURG. RES., vol. 35, 1983, pages 433 - 460
CHONG MS; HESTER J: "Diabetic painful neuropathy. Current and future treatment options", DRUGS, vol. 67, 2007, pages 569 - 585
COURTEIX C; ESCHALIER A; LAVARENNE J.: "Streptozocin-induced diabetic rats: behavioural evidence for a model of chronic pain", PDIN., vol. 53, 1993, pages 81 - 88
DYCK PJ; GIANNINI C.: "Pathologic alterations in the diabetic neuropathies of humans: a review", J.'NEUROPATHOL. EXP. NEURAL., vol. 55, 1996, pages 1181 - 1193
GREGG RW; MOLEPO JM; MONPETIT VJ; MIKAEL NZ; REDMOND D; GADIA M; STEWERT DJ.: "Cisplatin neurotoxicity: the relationship between dosage, time, and platinum concentration in neurologic tissues, and morphologic evidence of toxicity.", J CLIN. ONCOL., vol. 10, 1992, pages 795 - 803
JAKOBSEN J; LUNDBÆK K.: "Neuropathy in experimental diabetes: an animal model.", BR. MED. J., vol. 2, 1976, pages 278 - 279
MATTINGLY GE; FISCHER VW.: "Peripheral neuropathy following prolonged exposure to streptozotocin-induced diabetes in rats: a teased nerve fiber study.", ACTA NEUROPATHOL., vol. 59, 1983, pages 133 - 138
PERRY TA; HOLLOWAY HW; WEERASURIYA A; MOUTON PR; DUFFY K; MATTINSON JA; GREIG NH: "Evidence of GLP-1-mediated neuroprotection in an animal model of pyridoxine-induced peripheral sensory neuropathy", EXP. NEUROLOGY, vol. 203, 2007, pages 293 - 303
QUASTHOFF S; HARTUNG HP: "Chemotherapy-induced peripheral neuropathy.", J. NEUROL., vol. 249, 2002, pages 9 - 17
SCHMEICHEL AM; SCHMELZER JD; LOW PA: "Oxidative injury and apoptosis of dorsal root ganglion neurons in chronic experimental diabetic neuropathy.", DIABETES, vol. 52, 2003, pages 165 - 171
See also references of EP2212349A4 *
SZKUDELSKI T: "The mechanism of alloxan and streptozotocin action in B cells of the rat pancreas.", PHYSIOL. RES., vol. 50, 2001, pages 536 - 546
TOMLINSON DR; GARDINER NJ: "Glucose neurotoxicity.", NATURE NEUROSCI. REV., vol. 9, 2008, pages 36 - 45
WINDEBANK AJ; GRISOLD W.: "Chemotherapy-induced neuropathy.", J. PERIPHER. NERV. SYST., vol. 13, 2008, pages 27 - 46
WUARIN-BIERMAN L; ZAHND GR; KAUFMANN F; BURCKLEN L; ADLER J.: "Hyperalgesia in spontaneous and experimental animal models of diabetic neuropathy", DIABETOLOGIA., vol. 30, 1987, pages 653 - 658
YAGIHASHI S; YAMAGISHI S-I; WADA R.: "Pathology and pathogenic mechanisms of diabetic neuropathy: correlation with clinical signs and symptoms", DIAB. RES. CLIN. PRAC., vol. 77S, 2007, pages S 184 - S189

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012102625A2 (fr) * 2011-01-28 2012-08-02 Curonz Holdings Company Limited Composition thérapeutique
WO2012102625A3 (fr) * 2011-01-28 2012-11-01 Curonz Holdings Company Limited Composition thérapeutique
WO2012105854A2 (fr) * 2011-01-31 2012-08-09 Curonz Holdings Company Limited Compositions pour administration oculaire
WO2012105854A3 (fr) * 2011-01-31 2012-11-01 Curonz Holdings Company Limited Compositions pour administration oculaire
WO2014017927A1 (fr) 2012-07-27 2014-01-30 Curonz Holdings Company Limited Procédé de traitement d'une lésion du nerf optique, d'une ischémie ophtalmique ou d'une lésion de reperfusion ophtalmique
US20160015771A1 (en) * 2012-07-27 2016-01-21 Curonz Holdings Company Limited Method of treating optic nerve damage, ophthalmic ischemia or ophthalmic reperfusion injury
AU2013293645B2 (en) * 2012-07-27 2018-04-12 Curonz Holdings Company Limited Method of treating optic nerve damage, ophthalmic ischemia or ophthalmic reperfusion injury
EP2964332A4 (fr) * 2013-03-08 2016-09-14 Curonz Holdings Company Ltd Peptides de régénération neurale et leurs utilisations
WO2015084188A2 (fr) 2013-12-04 2015-06-11 Curonz Holdings Company Limited Méthode de traitement ou de prévention de l'épilepsie ou des crises d'épilepsie faisant appel à un ou plusieurs peptides stimulant la régénération neuronale
EP3076987A4 (fr) * 2013-12-04 2017-08-30 Curonz Holdings Company Limited Méthode de traitement ou de prévention de l'épilepsie ou des crises d'épilepsie faisant appel à un ou plusieurs peptides stimulant la régénération neuronale
AU2014357818B2 (en) * 2013-12-04 2020-06-11 Curonz Holdings Company Limited Method of treating or preventing epilepsy or seizures using one or more neural regeneration peptides

Also Published As

Publication number Publication date
NZ585363A (en) 2012-05-25
US20130231289A1 (en) 2013-09-05
US20110098228A1 (en) 2011-04-28
HRP20131016T1 (hr) 2014-01-03
US8309684B2 (en) 2012-11-13
EP2212349B1 (fr) 2013-08-07
EP2212349A4 (fr) 2010-09-29
US9650418B2 (en) 2017-05-16
PT2212349E (pt) 2013-11-07
AU2008314565A1 (en) 2009-04-23
DK2212349T3 (da) 2013-11-04
US9040485B2 (en) 2015-05-26
JP2011512322A (ja) 2011-04-21
JP5425086B2 (ja) 2014-02-26
ES2433568T3 (es) 2013-12-11
US20170218023A1 (en) 2017-08-03
EP2212349A1 (fr) 2010-08-04
PL2212349T3 (pl) 2014-01-31
CY1114619T1 (el) 2016-10-05
US20150307553A1 (en) 2015-10-29
AU2008314565B2 (en) 2013-12-12

Similar Documents

Publication Publication Date Title
US9650418B2 (en) Synthetic analogues of neural regeneration peptides
JP5555891B2 (ja) クモ毒から単離した新規ペプチド及びその使用
KR20240025721A (ko) 노화와 연관된 질환을 치료하기 위한 방법 및 조성물
CA2341810A1 (fr) Peptides cycliques derives de la prosaposine et leurs utilisations
JP2003503312A (ja) 立体選択的抗原線維形成ペプチドおよびそのペプチド模倣体
US10251937B2 (en) Retro-inverso analogs of spadin display increased antidepressant effects
EA015716B1 (ru) Применение sdf-1 для лечения и/или профилактики неврологических заболеваний
US8138304B2 (en) Neural regeneration peptides and formulations thereof
AU2010355825B9 (en) Treatment of vascular complications of diabetes
EP2382982A2 (fr) Peptides de régéneration neurale et procédés d'utilisation de ces derniers dans le traitement des lesions cerebrales
JP2002500161A (ja) 神経栄養性および鎮痛性のレトロインバースペプチド
KR20170104457A (ko) 신경변성 장애
WO2012015910A2 (fr) Utilisation de polypeptides adnf pour traitement de maladies neurodégénératives
WO1998042746A9 (fr) Peptides neurotrophiques de synthese derives de la saposine c
WO2024167821A2 (fr) Peptides pour le traitement de troubles neurologiques
CA2525107A1 (fr) Nouveaux facteurs de croissance des fibroblastes et procedes pour les utiliser

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08840727

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2010529981

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2008314565

Country of ref document: AU

Ref document number: 585363

Country of ref document: NZ

WWE Wipo information: entry into national phase

Ref document number: 2008840727

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2008314565

Country of ref document: AU

Date of ref document: 20081017

Kind code of ref document: A